Fluorescent protein-based imaging and tissue-specific RNA-seq analysis of Arabidopsis hydathodes
Fluorescent protein-based imaging and tissue-specific RNA-seq analysis of Arabidopsis hydathodes
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拟南芥基于荧光蛋白的成像和组织特异性 RNA-seq 分析
DOI：
10.1093/jxb/eraa519
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发表时间：
2020
影响因子：
6.9
通讯作者：
Shimada Tomoo
中科院分区：
文献类型：
--
作者：
Yagi Hiroki;Nagano Atsushi J;Kim Jaewook;Tamura Kentaro;Mochizuki Nobuyoshi;Nagatani Akira;Matsushita Tomonao;Shimada Tomoo
在维管植物的叶齿处通常发现有充水器，并参与将水分释放到外部。虽然已经在各种植物中进行了形态学和生理学的分析，但对参与包虫功能的基因知之甚少。在这项研究中，我们进行了基于荧光蛋白的成像和组织特异性RNA-seq分析在拟南芥hydathodes。我们使用了增强子捕获系E325，据报道，它在它的水囊表达绿色荧光蛋白（GFP）。我们发现E325-GFP在分布在气孔和木质部末端之间的小细胞（称为E细胞）中表达。韧皮部标记物pSUC 2：GFP和pSEOR 1：SEOR 1-YFP在根中未观察到荧光。这些观察结果表明，拟南芥的排水管是由三个主要组成部分：气孔，木质部末端，和E细胞。此外，我们使用E325-GFP系进行了包虫的转录组分析。用基于针的装置（直径约130 µm）从E325叶缘的GFP阳性或阴性区域收集微量样品。使用称为Lasy-Seq的高通量文库制备方法对每个单个微量样品进行RNA-seq。我们鉴定了72个差异表达基因。其中，68个基因显示出显着较高的表达和4个基因显示出显着较低的表达在包虫。我们的研究结果提供了新的见解的分子基础的水母的生理和发展。
Hydathodes are typically found at leaf teeth in vascular plants and are involved in water release to the outside. Although morphological and physiological analysis of hydathodes has been performed in various plants, little is known about the genes involved in hydathode function. In this study, we performed fluorescent protein-based imaging and tissue-specific RNA-seq analysis in Arabidopsis hydathodes. We used the enhancer trap line E325, which has been reported to express green fluorescent protein (GFP) at its hydathodes. We found that E325-GFP was expressed in small cells found inside the hydathodes (named E cells) that were distributed between the water pores and xylem ends. No fluorescence of the phloem markers pSUC2:GFP and pSEOR1:SEOR1-YFP was observed in the hydathodes. These observations indicate that Arabidopsis hydathodes are composed of three major components: water pores, xylem ends, and E cells. In addition, we performed transcriptome analysis of the hydathode using the E325-GFP line. Microsamples were collected from GFP-positive or -negative regions of E325 leaf margins with a needle-based device (~130 µm in diameter). RNA-seq was performed with each single microsample using a high-throughput library preparation method called Lasy-Seq. We identified 72 differentially expressed genes. Among them, 68 genes showed significantly higher and four genes showed significantly lower expression in the hydathode. Our results provide new insights into the molecular basis for hydathode physiology and development.